title: The YaaA protein of the Escherichia coli OxyR regulon lessens hydrogen peroxide toxicity by diminishing the amount of intracellular unincorporated iron.
abstract: Hydrogen peroxide (H(2)O(2)) is commonly formed in microbial habitats by either chemical oxidation processes or host defense responses. H(2)O(2) can penetrate membranes and damage key intracellular biomolecules, including DNA and iron-dependent enzymes. Bacteria defend themselves against this H(2)O(2) by inducing a regulon that engages multiple defensive strategies. A previous microarray study suggested that yaaA, an uncharacterized gene found in many bacteria, was induced by H(2)O(2) in Escherichia coli as part of its OxyR regulon. Here we confirm that yaaA is a key element of the stress response to H(2)O(2). In a catalase/peroxidase-deficient (Hpx(-)) background, yaaA deletion mutants grew poorly, filamented extensively, and lost substantial viability when they were cultured in aerobic LB medium. The results from a thyA forward mutagenesis assay and the growth defect of the yaaA deletion in a recombination-deficient (recA56) background indicated that yaaA mutants accumulated high levels of DNA damage. The growth defect of yaaA mutants could be suppressed by either the addition of iron chelators or mutations that slowed iron import, indicating that the DNA damage was caused by the Fenton reaction. Spin-trapping experiments confirmed that Hpx(-) yaaA cells had a higher hydroxyl radical (HO(•)) level. Electron paramagnetic resonance spectroscopy analysis showed that the proximate cause was an unusually high level of intracellular unincorporated iron. These results demonstrate that during periods of H(2)O(2) stress the induction of YaaA is a critical device to suppress intracellular iron levels; it thereby attenuates the Fenton reaction and the DNA damage that would otherwise result. The molecular mechanism of YaaA action remains unknown.
authors.affiliation: Department of Microbiology, University of Illinois at Urbana-Champaign, 601 South Goodwin Ave., Urbana, IL 61801, USA.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Bauer
authors.forename: James A
authors.initials: JA
authors.lastname: Imlay
mesh_terms: D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007501:Iron / Q000378:metabolism*; D008345:Manganese; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013312:Stress, Physiological / Q000187:drug effects
chemical_list: D029968:Escherichia coli Proteins; D012097:Repressor Proteins; C559319:YaaA protein, E coli; C499299:oxyR protein, E coli; D008345:Manganese; D006861:Hydrogen Peroxide; D007501:Iron